• International Journal of Stem Cells
• /
• 제16권3호
• /
• pp.342-355
• /
• 2023

Background and Objectives: Osteoblasts are derived from bone marrow mesenchymal stem cells (BMMSCs) and play important role in bone remodeling. While our previous studies have investigated the cell subtypes and heterogeneity in osteoblasts and BMMSCs separately, cell-to-cell communications between osteoblasts and BMMSCs in vivo in humans have not been characterized. The aim of this study was to investigate the cellular communication between human primary osteoblasts and bone marrow mesenchymal stem cells. Methods and Results: To investigate the cell-to-cell communications between osteoblasts and BMMSCs and identify new cell subtypes, we performed a systematic integration analysis with our single-cell RNA sequencing (scRNA-seq) transcriptomes data from BMMSCs and osteoblasts. We successfully identified a novel preosteoblasts subtype which highly expressed ATF3, CCL2, CXCL2 and IRF1. Biological functional annotations of the transcriptomes suggested that the novel preosteoblasts subtype may inhibit osteoblasts differentiation, maintain cells to a less differentiated status and recruit osteoclasts. Ligand-receptor interaction analysis showed strong interaction between mature osteoblasts and BMMSCs. Meanwhile, we found FZD1 was highly expressed in BMMSCs of osteogenic differentiation direction. WIF1 and SFRP4, which were highly expressed in mature osteoblasts were reported to inhibit osteogenic differentiation. We speculated that WIF1 and sFRP4 expressed in mature osteoblasts inhibited the binding of FZD1 to Wnt ligand in BMMSCs, thereby further inhibiting osteogenic differentiation of BMMSCs. Conclusions: Our study provided a more systematic and comprehensive understanding of the heterogeneity of osteogenic cells. At the single cell level, this study provided insights into the cell-to-cell communications between BMMSCs and osteoblasts and mature osteoblasts may mediate negative feedback regulation of osteogenesis process.

• Asian-Australasian Journal of Animal Sciences
• /
• 제28권9호
• /
• pp.1235-1243
• /
• 2015

The goat Capra hircus is one of several economically important livestock in China. Advances in molecular genetics have led to the identification of several single nucleotide variation markers associated with genes affecting economic traits. Validation of single nucleotide variations in a whole-transcriptome sequencing is critical for understanding the information of molecular genetics. In this paper, we aim to develop a large amount of convinced single nucleotide polymorphisms (SNPs) for Cashmere goat through transcriptome sequencing. In this study, the transcriptomes of Cashmere goat skin at four stages were measured using RNA-sequencing and 90% to 92% unique-mapped-reads were obtained from total-mapped-reads. A total of 56,231 putative SNPs distributed among 10,057 genes were identified. The average minor allele frequency of total SNPs was 18%. GO and KEGG pathway analysis were conducted to analyze the genes containing SNPs. Our follow up biological validation revealed that 64% of SNPs were true SNPs. Our results show that RNA-sequencing is a fast and efficient method for identification of a large number of SNPs. This work provides significant genetic resources for further research on Cashmere goats, especially for the high density linkage map construction and genome-wide association studies.

• Journal of Microbiology and Biotechnology
• /
• 제26권7호
• /
• pp.1242-1251
• /
• 2016

The ethanol production of Trichoderma reesei was improved by genome shuffling in our previous work. Using RNA-Seq, the transcriptomes of T. reesei wild-type CICC40360 and recombinant strain HJ48 were compared under fermentation conditions. Based on this analysis, we defined a set of T. reesei genes involved in ethanol production. Further expression analysis identified a series of glycolysis enzymes, which are upregulated in the recombinant strain HJ48 under fermentation conditions. The differentially expressed genes were further validated by qPCR. The present study will be helpful for future studies on ethanol fermentation as well as the roles of the involved genes. This research reveals several major differences in metabolic pathways between recombinant strain HJ48 and wild-type CICC40360, which relates to the higher ethanol production on the former, and their further research could promote the development of techniques for increasing ethanol production.

• BMB Reports
• /
• 제48권8호
• /
• pp.427-428
• /
• 2015

Despite high interests on microenvironmental regulation of leukemic cells, little is known for bone marrow (BM) niche in leukemia patients. Our recent study on BMs of acute myeloid leukemia (AML) patients showed that the mesenchymal stromal cells (MSCs) are altered during leukemic conditions in a clinical course-dependent manner. Leukemic blasts caused reprogramming of transcriptomes in MSCs and remodeling of niche cross-talk, selectively suppressing normal primitive hematopoietic cells while supporting leukemogenesis and chemo-resistance. Notably, differences in BM stromal remodeling were correlated to heterogeneity in subsequent clinical courses of AML, i.e., low numbers of mesenchymal progenitors at initial diagnosis were correlated to complete remission for 5-8 years, and high contents of mesenchymal progenitor or MSCs correlated to early or late relapse, respectively. Thus, stromal remodeling by leukemic cell is an intrinsic part of leukemogenesis that can contribute to the clonal dominance of leukemic cells over normal hematopoietic cells, and can serve as a biomarker for prediction of prognosis. [BMB Reports 2015; 48(8): 427-428]

• ALGAE
• /
• 제30권4호
• /
• pp.303-312
• /
• 2015

The intimate physical interaction between food algae and sacoglossan sea slug is a pertinent system to test the theory that “you are what you eat.” Some sacoglossan mollusks ingest and maintain chloroplasts that they acquire from the algae for photosynthesis. The basis of photosynthesis maintenance in these sea slugs was often explained by extensive horizontal gene transfer (HGT) from the food algae to the animal nucleus. Two large-scale expressed sequence tags databases of the green alga Bryopsis plumosa and sea slug Placida dendritica were established using 454 pyrosequencing. Comparison of the transcriptomes showed no possible case of putative HGT, except an actin gene from P. dendritica, designated as PdActin04, which showed 98.9% identity in DNA sequence with the complementary gene from B. plumosa, BpActin03. Highly conserved homologues of this actin gene were found from related green algae, but not in other photosynthetic sea slugs. Phylogenetic analysis showed incongruence between the gene and known organismal phylogenies of the two species. Our data suggest that HGT is not the primary reason underlying the maintenance of short-term kleptoplastidy in Placida dendritica.

• ALGAE
• /
• 제33권2호
• /
• pp.181-189
• /
• 2018

Emiliania huxleyi (a haptophyte) is the most abundant coccolithophore species that produces delicate calcite scales called coccoliths. In this study, we identified several candidate genes associated with coccolith production by comparing the transcriptomes of the calcifying (CCMP 371) and non-calcifying (CCMP 2090) strains of E. huxleyi. Among the candidates, genes highly expressed in CCMP 371 were identified. To confirm whether these genes are associated with calcification, we modulated coccolith production in CCMP 371 by culturing it at different calcium concentrations. At an ambient (10 mM) concentration of calcium in the growth medium, CCMP 371 sustained its calcifying ability. However, at a low (0.1 mM) concentration or absence of calcium, there was no calcite formation, demonstrating that calcium-limiting conditions negatively affect calcification. We also evaluated the expression patterns of the putative genes in cells grown at different calcium concentrations by quantitative reverse transcription polymerase chain reaction. In addition, we showed that the growth rate of cells cultured under calcium-limiting conditions does not differ from that under ambient conditions. Further studies are required to investigate the roles of the putative calcification-associated genes at the molecular level.

• Clinical and Experimental Reproductive Medicine
• /
• 제39권3호
• /
• pp.95-106
• /
• 2012

It has been revealed that multiple cohorts of tertiary follicles develop during some animal estrous cycle and the human menstrual cycle. To reach developmental competence, oocytes need the support of somatic cells. During embryogenesis, the primordial germ cells appear, travel to the gonadal rudiments, and form follicles. The female germ cells develop within the somatic cells of the ovary, granulosa cells, and theca cells. How the oocyte and follicle cells support each other has been seriously studied. The latest technologies in genes and proteins and genetic engineering have allowed us to collect a great deal of information about folliculogenesis. For example, a few web pages (http://www.ncbi.nlm. nih.gov; http://mrg.genetics.washington.edu) provide access to databases of genomes, sequences of transcriptomes, and various tools for analyzing and discovering genes important in ovarian development. Formation of the antrum (tertiary follicle) is the final phase of folliculogenesis and the transition from intraovarian to extraovian regulation. This final step coordinates with the hypothalamic-pituitary-ovarian axis. On the other hand, currently, follicle physiology is under intense investigation, as little is known about how to overcome women's ovarian problems or how to develop competent oocytes from in vitro follicle culture or transplantation. In this review, some of the known roles of hormones and some of the genes involved in tertiary follicle growth and the general characteristics of tertiary follicles are summarized. In addition, in vitro culture of tertiary follicles is also discussed as a study model and an assisted reproductive technology model.

• Journal of Plant Biotechnology
• /
• 제43권2호
• /
• pp.189-203
• /
• 2016

Brassica oleraceae var capitata is a member of the Brassicaceae family and is widely used as an horticultural crop. In the present study, transcriptome analysis of B. oleraceae L. var capitata was done for the first time using eight-week old seedlings treated with $50{\mu}m$ MeJA, versus mock-treated samples. The complete transcripts for both samples were obtained using the GS-FLX sequencer. Overall, we obtained 275,570 and 266,457 reads from seedlings treated with or without $50{\mu}m$ MeJA, respectively. All the obtained reads were annotated using biological databases and functionally classified using gene ontology (GO), the Kyoto Encyclopedia of Genes and Genomics (KEGG). By using GO analyses, putative transcripts were examined in terms of biotic and abiotic stresses, cellular component organization, biogenesis, and secondary metabolic processes. The KEGG pathways for most of the transcripts were involved in carbohydrate metabolism, energy metabolism, and secondary metabolite synthesis. In order to double the sequenced data, we randomly chose two putative genes involved in terpene biosynthetic pathways and studied their transcript patterns under MeJA treatment. This study will provide us a platform to further characterize the genes in B. oleracea var capitata.

• Genomics & Informatics
• /
• 제17권3호
• /
• pp.27.1-27.9
• /
• 2019

Supernumerary B chromosomes were found in Lilium amabile (2n = 2x = 24), an endemic Korean lily that grows in the wild throughout the Korean Peninsula. The extra B chromosomes do not affect the host-plant morphology; therefore, whole transcriptome analysis was performed in 0B and 1B plants to identify differentially expressed genes. A total of 154,810 transcripts were obtained from over 10 Gbp data by de novo assembly. By mapping the raw reads to the de novo transcripts, we identified 7,852 differentially expressed genes (log₂FC > |10|), in which 4,059 and 3,794 were up-and down-regulated, respectively, in 1B plants compared to 0B plants. Functional enrichment analysis revealed that various differentially expressed genes were involved in cellular processes including the cell cycle, chromosome breakage and repair, and microtubule formation; all of which may be related to the occurrence and maintenance of B chromosomes. Our data provide insight into transcriptomic changes and evolution of plant B chromosomes and deliver an informative database for future study of B chromosome transcriptomes in the Korean lily.

• 식물분류학회지
• /
• 제50권1호
• /
• pp.22-26
• /
• 2020

Chrysosplenium aureobracteatum Y. I. Kim & Y. D. Kim (Saxifragaceae) is a recently described endemic species growing in the central part of the Korean peninsula. It requires constant monitoring for conservation due to its limited distributions. There is also a need for molecular markers for proper assessments of the genetic differentiation of C. aureobracteatum from species morphologically similar to it. In this study, we developed microsatellite markers that can be used to evaluate the genetic diversity of this species, representing fundamental data with which to conserve the natural populations of the species. A total of 17 expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed by the Illumina pair-end sequencing of the transcriptomes of C. aureobracteatum. These markers were successfully applied to populations of C. aureobracteatum and to its most closely related species, C. barbatum, revealing high polymorphism in both species. The EST-SSR markers developed in this study were proven to be useful not only to monitor the population genetic structure of C. aureobracteatum for conservation purposes but also to study the genetic delimitation of the species from species closely related to it.